ABSTRACT This program aims to optimize a method for banking clinical-grade bone marrow from organ donors that can be deployed nationally to ameliorate shortages, provide bone marrow stockpiles as medical countermeasures for nuclear threats, and support existing research approaches such as delayed immune tolerance induction and transfusion of HIV-resistant stem cells. In Phase I we adapted protocols developed by our research team for procurement, processing, and banking bone marrow from cadaveric vertebral bodies and ilia, which were shown to produce high yields of viable hematopoietic stem cells and other bone marrow cells from donors after varying lengths of warm and cold ischemic times. Equations have been derived from these data which have allowed modeling the effects of warm and cold ischemia on hematopoietic stem cell viability and function. These predictive models provide the tools necessary for completing optimization of the recovery and isolation protocols. In this proposal we will make key refinements to our method for deployment on a large scale, which will allow recovery at geographically diverse organ procurement agencies and shipment to a centralized facility for processing. The end result will be a large supply of bone marrow that can be used on-demand, off the shelf, complementing existing bone marrow donor registries and cord blood banks. The technical objectives of this Phase 2 study are to (1) optimize preservation of stem cells in whole vertebrae shipped to Ossium?s central processing facility, (2) maximize recovery of viable stem cells exposed to long warm ischemia times, and (3) validate the final optimized process. In Aim 1, we will optimize preservation solutions for procurement and shipment of donor vertebral body bone marrow. In Aim 2, we will optimize whole bone marrow isolation solutions to maximize recovery of viable cells. In Aim 3, we will validate all processes by building and testing a pilot bank and comparing deceased donor and aspirated bone marrow in a mouse xenotransplantation study. The outcome of these aims will determine the most effective system for procurement, processing and banking of cadaveric bone marrow for clinical use on a large scale.